Charland, Nathalie

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Engineering Conferences InternationalECI Digital ArchivesVaccine Technology IV ProceedingsSummer 5-24-2012Plant-Made Influenza Virus-Like Particles: forPandemic and BeyondNathalie CharlandMedicagoFollow this and additional works at: http://dc.engconfintl.org/vaccine_ivPart of the Biomedical Engineering and Bioengineering CommonsThis Conference Proceeding is brought to you for free and open access by the Proceedings at ECI Digital Archives. It has been accepted for inclusion inVaccine Technology IV by an authorized administrator of ECI Digital Archives. For more information, please contact franco@bepress.com.Recommended CitationNathalie Charland, "Plant-Made Influenza Virus-Like Particles: for Pandemic and Beyond" in "Vaccine Technology IV", B. Buckland,University College London, UK; J. Aunins, Janis Biologics, LLC; P. Alves , ITQB/IBET; K. Jansen, Wyeth Vaccine Research Eds, ECISymposium Series, (2013). http://dc.engconfintl.org/vaccine_iv/44Plant-Made Influenza Virus-Like Particles: for Pandemic and Beyond© 2012 Medicago Inc.All rights reservedNathalie Charland, PhDDirector, Product PortfolioVaccine Technology IVAlbufeira, PortugalMay  24, 20122Forward-Looking Statements and Disclaimers• This presentation contains forward-looking information and statements which constitute “forward-looking information” under Canadian securities law and which may be material regarding, among other things, the Company’s beliefs, plans, objectives, estimates, intentions and expectations. Forward-looking information and statements are typically identified by words such as “anticipate”, “believe”, “expect”, “estimate”, “forecast”, “goal”, “intend”, “plan”, “will”, “may”, “should”, “could” and similar expressions. Specific forward-looking information in this document includes, but is not limited to, statements with respect to the Company’s future operating and financial results, its research and development activities, its capital expenditure plans and the ability to execute on its future operating, investing and financing strategies.  These forward-looking information and statements, by their nature, necessarily involve risks and uncertainties that could cause actual results to differ materially from those contemplated by these forward-looking statements. We consider the assumptions on which these forward-looking statements are based to be reasonable, but caution the reader that these assumptions regarding future events, many of which are beyond our control, may ultimately prove to be incorrect since they are © 2012 Medicago Inc.All rights reservedsubject to risks and uncertainties that affect us.  Additional information regarding risk factors can be found in the Medicago Annual Information Form filed on March 29, 2012.• Our statements of “belief” in respect of our product and partner product candidates are basedprimarily upon our results derived to date from our research and development program. We believethat we have a reasonable scientific basis upon which we have made such statements. It is notpossible, however, to predict, based upon studies in vitro and animal studies whether a newtherapeutic agent or technology will be proved to be safe and/or effective in humans. We cannotassure that the particular results expected by us will occur.• Any forward-looking statements and statements of “belief” represent our estimates only andshould not be relied upon as representing our estimates as of any subsequent date. Except asrequired by law, we do not assume any obligation to update any forward looking statements orstatements of “belief”. We disclaim any intention or obligation to update or revise any forward-looking statements or statements of “belief”, whether as a result of new information, futureevents or otherwise. Nothing herein should be construed as an Offering of securities of theCompany in any jurisdictions.3Reengineering vaccine design and production (beyond pandemic)• Strains with pandemic potential are circulating every year; experts agree only a matter of time before a highly lethal strain develops due to strain mutations– Kawaoka’s & Fouchier’s studies• Rapid response in pandemic vaccine production is needed to © 2012 Medicago Inc.All rights reservedaddress rapid rates of infection & minimize deaths– In 2009 pandemic, time required to manufacture both egg- & cell-based vaccines resulted in “too little, too late” vaccine responses  no significant impact*• Need for flexible technologies to respond to emerging or engineered biothreats– Multi-product platform*Ref: Osterholm & Kelley (2012) Mammalian-transmissible H5N1 influenza: Facts and Perspective. mBio 3(2) doi:10.1128/mBio.00045-124Medicago overviewFocus Vaccines & Protein-based pharmaceuticalsManufacturing technology Transient expression in TobaccoVaccine technology Virus-like particlesVaccine discovery platform VLPExpressHeadquarters, laboratories & Quebec City, CANADA© 2012 Medicago Inc.All rights reservedcGMP facilitiesResearch Triangle Park, NC, USAGenopole d’Evry, FRANCEAgreementsMitsubishi Tanabe Pharma – Strategic AllianceDARPA  Award – US$21M IDRI – Phase I H5 GLA, intradermalPharma & U.S. Army – Vaccines outside of flu$35M & 12 months to buildCapacity of 10M doses/month5Preclinical Phase 1 Phase 2 Phase 3Pandemic influenzaH5H5 – Intradermal+GLAH1Products© 2012 Medicago Inc.All rights reservedSeasonal influenzaQuadrivalentRabiesUndisclosedEbolaRotavirusTop 10 Pharma6Protalix is paving the way…First plant-derived pharmaceutical approved by FDA on May 1st, 2012© 2012 Medicago Inc.All rights reserved7Medicago’s manufacturing technologyProprietary platform for the production recombinant proteins based on a transient expression technology in plants (N. benthamiana)Plants & Agrobacterium © 2012 Medicago Inc.All rights reservedInfiltrationPurificationIncubationMedicago VLP Influenza virusExtraction• 19 days to 1st lot• Rapid lot-release:• HA content• 14-days sterility testpreparation8Characteristics of the plant-made H5 VLP vaccine• Lot-release assays include purity, potency by SRID, lipid and proteincontent, residual DNA, endotoxins, nicotine and anabasine content, sterility• Additional characterization such as SEC, NTA, MFI, PDI, glycans• Product purity 98%  (HA0, HA1, HA2) Silver stained SDS PAGECoomassie stained SDS PAGEWesternblot© 2012 Medicago Inc.All rights reserved2251501007550352515KDaHA0HA1HA2HA dimers 2251501007550352515KDaHA0HA1HA2HA dimers9General safety profilePhase 1 H1 VLP and Phase 2 part A H5 VLP10203040506070% of incidence© 2012 Medicago Inc.All rights reserved0Non-adjuvanted H1 VLPPlacebo Phase 1Adjuvanted H5 VLPNon-adjuvanted H5 VLPPlacebo Phase 2 part ANon-adjuvanted H1 VLPPlacebo Phase 1Adjuvanted H5 VLPNon-adjuvanted H5 VLPPlacebo Phase 2 part ANon-adjuvanted H1 VLPPlacebo Phase 1Adjuvanted H5 VLPNon-adjuvanted H5 VLPPlacebo Phase 2 part ARedness Swelling Pain% of incidenceSolicited local AEsSevereModerateMildSimilar profiles were observed for systemic AEs10SafetyIgE to plant glycansClinical trial Group Number of subjects with IgEs ≥grade 1 to bromelain at screening Number of subjects that showed an IgEincrease after vaccinationNumber of subjects that showed detectable IgEs6 months after vaccinationPhase 1 with H1 VLP (one dose)Non-adjuvanted VLP (n=58)3.5%(2/57)0%(0/57)1.8%(1/56)Fluzone 0% 0% 0%© 2012 Medicago Inc.All rights reserved(trivalent, n=20) (0/20) (0/20) (0/20)Placebo (n=20) 0%(0/20)0%(0/20)0%(0/20)Phase 2 with H5 VLP (two doses)Adjuvanted VLP (n=192)3%(6/191)0%(0/188)3%(6/191)Non-adjuvanted VLP (n=29)7%(2/29)0%(0/29)4%(1/27)Placebo (n=27) 0%(0/28)0%(0/28)0%(0/28)• No onset of allergic reactions correlating with in vitro assay(manuscript in preparation)11Antibody response – H5 VLPHI testH5 VLP Vaccine5 µg +Al 10 µg +Al20 µg +Al30 µg +Al45 µg +Al45 µg Seroconversion rate after two doses of H5 VLP (18-64 years of age)© 2012 Medicago Inc.All rights reservedPhase I*(groups of 12 subj.)16.7(0.02-0.48)25.0(0.06-0.57)58.3(0.28-0.85)Phase II (part A, groups of 30 subj)58.6(0.39-0.77)53.6(0.34-0.73)46.7(0.28-0.66)21.4(0.08-0.41)*12Antibody response – H5 VLP Phase II trial part BSeroconversion rates after two dosesH5 VLP Vaccine, 20 µg dose + alum18-29 n=2330-39n=2240-49n=2650-60n=26© 2012 Medicago Inc.All rights reservedHI test 56.5% 45.5% 38.5% 50.0%SRH test 52.2% 40.9% 50.0% 42.3%MN test 52.2% 50.0% 50.0% 50.0%All vaccine doses statistically different from placeboH5 VLP vaccine equally immunogenic in young and older adults13Duration of antibody response – H5 VLPHI titers for homologous strainDose of H5 VLP (µg)% of subjects with positive antibody response (n = 15)Pre-vaccinationAfter 1stdoseAfter 2nddose6-month after boost© 2012 Medicago Inc.All rights reserved20 + alum 0 23,3 75,9 69,030 + alum 0 24,1 75,0 50,045 + alum 0 23,3 73,3 60,045 0 10,7 64,3 40,7Placebo 0 0 0 8,0Alum showed benefit on the antibody level for the H5 VLP vaccine although formulation not optimized at high vaccine dosages14H1 VLP Vaccine (groups of 20 subj.)5 µg 13 µg 28 µgAntibody response – H1 VLPHI testSeroconversion rate after one single non-adjuvanted dose of H1 VLP (18-49 years of age)© 2012 Medicago Inc.All rights reserved61.1(0.36-0.83)63.2(0.38-0.84)82.4(0.60-0.97)15Memory CD8+ T CellsMemory CD4+ T CellsH1 (VLP) ResponseCD8+ T cellsCD4+ T cellsH1 VLP Vaccine Induced a Long Lasting Memory Multifunctional T cell Response© 2012 Medicago Inc.All rights reservedH1 (VLP) ResponsePer 106CD45RA-CD8+ T cellsH1 (VLP) ResponsePer 106CD45RA-CD4+ T cellsPolyfunctional T cells16H1 VLP Vaccine Induces Cross-Reactive CD4+ T cell Responses against HA (H5N1)Total CD4+ T cell response against HA (H1N1) :• PBMC at +6 month post-vaccine• In vitro stimulation with H5 VLP and cytokine detectionResponding Cells T cells© 2012 Medicago Inc.All rights reserved* P<0.05 (Mann-Whitney)Responding Cells per 106CD4+T cellsPlacebo H1 VLPSimilar results obtained for CD8+ T cell responses17Main conclusions from clinical trials• Safety– Safe and well tolerated– More than 400 subjects dosed– 1 or 2 doses– With or without alum– Dosages up to 45 µg© 2012 Medicago Inc.All rights reserved– No onset of allergic reactions• Immunogenicity– The HA-VLP vaccine induces durable antibody response– Durable, poly-functional and cross-reactive T Cell responses to influenza HA antigens– Strong innate immune response induced (data not shown)18• Can the manufacturing and vaccine platforms also produce VLPs presenting non-influenza antigens?– different genomic organization (RNA, DNA, polyprotein…)– different envelope proteins structure– different budding sites and requirementsExpanding the applications of the manufacturing and vaccine platforms© 2012 Medicago Inc.All rights reservedHIVRSVInfluenzaHepatitis C Human papillomavirus19VLPExpress: Accelerating the discovery of VLP-based vaccines• Scale-down of manufacturing technology– HT cloning (96 DNA constructs in 10 days)– Automated HT infiltration (50 expression strategies/days)© 2012 Medicago Inc.All rights reserved– HT monitoring of viral protein and VLP accumulation– Generic purification method• Screening – Capacity improved 10X – Time to identification 10X faster20Flexibility: Broadening applications of manufacturing & vaccine platforms• Throughput of VLPExpress enables running multiple projects in parallel• In 2011, the discovery team has tested >2,000 engineering approaches and identified those driving the assembly of VLPs for 7 different families of viruses:– Influenza (including 20 sub-types)– Rabies– Proprietary undisclosed  enveloped VLP© 2012 Medicago Inc.All rights reserved– Ebolavirus– Varicella-zoster virus– HIV– Partnered undisclosed capsid VLP• Advantages– Testing of multiple expression vectors in parallel– Comparing different protein engineering strategies– Co-expressing various proteins using separate Agrobacteria– Rapid optimization of expression conditions– Allow development of vaccines with "built-in" efficacy and manufacturing readiness21Advantages of Medicago’s plant-based technology• Pandemic– Target any HA sequence including wild-type– Induce strong antibody responses– Induce a balanced immune response• Strong CD4+ and CD8+ memory• Cross-reactive for other strains© 2012 Medicago Inc.All rights reserved– More durable strain-specific immunity– Superior cross-reactivity– Possible role in the elderly• Other emerging threats– Many features of plant-made influenza VLPs vaccines applicable to other indications (natural or man-made)– Flexibility, speed, scale-up & cost advantages22AcknowledgementsAt MedicagoResearch and Innovation teamProduct Development teamManufacturing and QA teams CollaboratorsGLA adjuvanted vaccineFrom IDRI: Darrick Carter Patti Hon        Anthony DesbienSteve Reed Rhea Coler© 2012 Medicago Inc.All rights reservedSponsorsSusan Baldwin Chris FoxFrom CSU (animal studies): Richard BowenCell-mediated Immune responseImmuneCarta: David FavreMcGill University: Brian J. Ward’s teamObrigada!© 2012 Medicago Inc.All rights reservedTSX: MDGwww.medicago.com

Plant-Made Influenza Virus-Like Particles: for Pandemic and Beyond

https://dc.engconfintl.org/cgi/viewcontent.cgi?article=1042&amp;context=vaccine_iv

Plant-Made Influenza Virus-Like Particles: for Pandemic and Beyond

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